US7298550B2 - Dichroic mirror, fluorescence filter set, and fluoroscopy apparatus - Google Patents

Dichroic mirror, fluorescence filter set, and fluoroscopy apparatus Download PDF

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US7298550B2
US7298550B2 US11/184,814 US18481405A US7298550B2 US 7298550 B2 US7298550 B2 US 7298550B2 US 18481405 A US18481405 A US 18481405A US 7298550 B2 US7298550 B2 US 7298550B2
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wavelength
dichroic mirror
refractive
band
fluorescence
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US20060028729A1 (en
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Daisuke Nishiwaki
Kei Kikuchi
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Evident Corp
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Olympus Corp
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/645—Specially adapted constructive features of fluorimeters
    • G01N21/6456—Spatial resolved fluorescence measurements; Imaging
    • G01N21/6458—Fluorescence microscopy
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/16—Microscopes adapted for ultraviolet illumination ; Fluorescence microscopes
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B27/00—Optical systems or apparatus not provided for by any of the groups G02B1/00 - G02B26/00, G02B30/00
    • G02B27/10—Beam splitting or combining systems
    • G02B27/1006—Beam splitting or combining systems for splitting or combining different wavelengths
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B27/00—Optical systems or apparatus not provided for by any of the groups G02B1/00 - G02B26/00, G02B30/00
    • G02B27/10—Beam splitting or combining systems
    • G02B27/14—Beam splitting or combining systems operating by reflection only
    • G02B27/141—Beam splitting or combining systems operating by reflection only using dichroic mirrors
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B27/00—Optical systems or apparatus not provided for by any of the groups G02B1/00 - G02B26/00, G02B30/00
    • G02B27/10—Beam splitting or combining systems
    • G02B27/14—Beam splitting or combining systems operating by reflection only
    • G02B27/142—Coating structures, e.g. thin films multilayers
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B5/00—Optical elements other than lenses
    • G02B5/20—Filters
    • G02B5/28—Interference filters
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B5/00—Optical elements other than lenses
    • G02B5/20—Filters
    • G02B5/28—Interference filters
    • G02B5/285—Interference filters comprising deposited thin solid films
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/645—Specially adapted constructive features of fluorimeters
    • G01N2021/6463—Optics
    • G01N2021/6471—Special filters, filter wheel

Definitions

  • the present invention relates to a dichroic mirror, to a fluorescence filter set, and to a fluoroscopy apparatus, such as a microscope or endoscope for fluoroscopy, a measurement device for measuring fluorescence intensity, and so forth.
  • a fluoroscopy apparatus such as a microscope or endoscope for fluoroscopy, a measurement device for measuring fluorescence intensity, and so forth.
  • Known dichroic mirrors according to the related art include those disclosed below, for example.
  • a dichroic mirror described in Japanese Unexamined Patent Application Publication No. HEI-11-202127 includes a multilayer film formed, on a transparent substrate, by alternately laminating high-refractive-index layers, intermediate-refractive-index layers, and low-refractive-index layers having optical film thicknesses equal to 1 ⁇ 4 of a design center wavelength ⁇ 0 .
  • a dichroic mirror described in Japanese Unexamined Patent Application Publication No. SHO-61-45202 includes alternating layers formed, on a substrate, by alternately laminating high-refractive-index layers having an optical film thickness equal to 1 ⁇ 4 of a reference wavelength ⁇ 0 , and low-refractive-index layers having an optical film thickness equal to 3 ⁇ 4 of the reference wavelength ⁇ 0 .
  • a dichroic mirror described in Japanese Unexamined Patent Application Publication No. SHO-54-110855 includes alternating layers formed by alternately laminating, on a substrate, high-refractive-index layers having optical film thicknesses equal to 1 ⁇ 4 and 3 ⁇ 4 of a reference wavelength ⁇ 0 , and low-refractive-index layers having optical film thicknesses equal to 1 ⁇ 4 and 3 ⁇ 4 of the reference wavelength ⁇ 0 .
  • One effective method in fluoroscopy is to set a reflection band for the excitation light and a transmission band for the fluorescence of the dichroic mirror as close to each other as possible, because the wavelength of excitation light irradiating a specimen and the wavelength of fluorescence produced by a fluorescent substance in the specimen are close to each other.
  • the dichroic mirror described in Japanese Unexamined Patent Application Publication No. HEI-11-202127 suffers from the drawback that, if the dichroic mirror is placed at an angle of about 45° with respect to the incident optical axis of the excitation light, a relatively large step occurs in the transmission characteristic due to splitting of P-polarized and S-polarized light.
  • the reflection band for excitation light an d the transmission band for fluorescence are separated by a distance corresponding to the width of the region where the step is formed.
  • the transmission characteristic is set at the short-wavelength side in order to efficiently collect weak fluorescence emitted from the specimen, part of the excitation light will be transmitted by the dichroic mirror, whereas if the transmission characteristic is set at the long-wavelength side in order to completely reflect the excitation light, part of the fluorescence will be reflected by the dichroic mirror.
  • the present invention has been conceived in light of the circumstances described above, and an object thereof is to provide a dichroic mirror having a transmission characteristic in which a step due to splitting of P-polarized and S-polarized light can be reduced and having a fluorescence-transmitting band extending to long wavelengths.
  • Another object is to provide a fluorescence filter set that can be constructed of environmentally conscious glass materials that do no use hazardous substances, such as lead, and a fluoroscopy apparatus including such a fluorescence filter set, such as a microscope or endoscope, or a measurement device for measuring fluorescence intensity.
  • the present invention provides the following solutions.
  • the present invention provides a dichroic mirror including a transparent substrate; and a dielectric multilayer film formed by alternately laminating high-refractive-index layers and low-refractive-index layers on the transparent substrate.
  • the dielectric multilayer film includes from 50 to 150 alternately laminated high-refractive-index layers and low-refractive-index layers each having an optical film thickness from 1.5 ⁇ 0 /4 to 2.5 ⁇ 0 /4, where ⁇ 0 is a design wavelength, and wherein the dichroic mirror has a reflection band located at wavelengths shorter than the design wavelength.
  • the width of a higher-harmonic reflection band at or above the second harmonic can be increased and used as the reflection band.
  • the higher-harmonic reflection band even if the dichroic mirror is tilted with respect to the incident optical axis of the excitation light, a step in the transmission characteristic due to splitting of P-polarized and S-polarized light components can be reduced, which in turn allows the reflection band for the excitation light and the transmission band for the fluorescence to be made close to each other. As a result, substantially all of the excitation light can be reflected and the fluorescence can be efficiently transmitted.
  • a fluorescence-transmitting band extending to longer wavelengths can be ensured, to allow a wider range of use, even for collecting long-wavelength fluorescence.
  • the present invention also provides a fluorescence filter set including a first wavelength-selecting member for selectively transmitting excitation light of a first wavelength; a dichroic mirror described above, for reflecting the excitation light transmitted by the first wavelength-selecting member in a direction orthogonal to an incident direction and for transmitting fluorescence returning from the reflection direction; and a second wavelength-selecting member for selectively transmitting the fluorescence transmitted by the dichroic mirror.
  • the fluorescence filter set when the excitation light selectively transmitted by the first wavelength-selecting member is incident on the dichroic mirror, it is reflected in a direction orthogonal to the incident direction and is irradiated in the reflection direction.
  • the fluorescence returning from the reflection direction is incident on the dichroic mirror, it is transmitted thereby and is incident on the second wavelength-selecting member placed at the subsequent position, which selectively transmits only the fluorescence.
  • the fluorescence filter set can be constructed using only environmentally conscious glass materials that do not include harmful substances, such as lead or cadmium.
  • the present invention provides a fluoroscopy apparatus, such as a microscope or endoscope or a measurement device for measuring fluorescence intensity, including: a light source; a fluorescence filter set described above, for reflecting excitation light from the light source and for transmitting fluorescence returning from the reflection direction; an objective optical system for irradiating a specimen with the excitation light reflected by the fluorescence filter set; and an observation optical system for observing the fluorescence returning from the specimen and transmitted by the objective optical system and the fluorescence filter set.
  • a fluoroscopy apparatus such as a microscope or endoscope or a measurement device for measuring fluorescence intensity
  • a fluorescence filter set described above, for reflecting excitation light from the light source and for transmitting fluorescence returning from the reflection direction
  • an objective optical system for irradiating a specimen with the excitation light reflected by the fluorescence filter set
  • an observation optical system for observing the fluorescence returning from the specimen and transmitted by the objective optical system and the fluorescence filter set.
  • the fluoroscopy apparatus when light emitted from the light source is incident on the fluorescence filter set, the excitation light selectively transmitted by the first wavelength-selecting member is reflected by the dichroic mirror and is irradiated onto the specimen via the objective optical system.
  • a fluorescent substance injected in advance or an autofluorescence substance in the specimen is excited by the excitation light and fluorescence is produced.
  • the fluorescence produced in the specimen returns via the objective optical system, enters the fluorescence filter set, is transmitted by the dichroic mirror, passes through the second wavelength-selecting member, and is introduced to the observation optical system.
  • the fluoroscopy apparatus since substantially all of the excitation light from the light source is reflected by the dichroic mirror, no excitation light is incident on the second wavelength-selecting member from various directions, and therefore, it is possible to eliminate substantially all of the excitation light incident on the observation optical system, without using colored glass in the second wavelength-selecting member.
  • the glass members of the fluoroscopy apparatus can be constructed using only environmentally conscious glass materials that do not include harmful substances, and it is therefore possible to construct a fluoroscopy apparatus that is suitable from the viewpoint of environmental protection. Also, it is possible to almost completely prevent the excitation light from entering the observation optical system, even without using colored glass elements, thus allowing clear, flare-free observation images to be obtained.
  • the step in the transmission characteristic due to splitting of P-polarized and S-polarized light can be reduced, and a fluorescence-transmitting band extending to long wavelengths can be ensured.
  • the fluorescence filter set and the fluoroscopy apparatus according to the aspects of the invention described above the excitation light is not transmitted to the observation optical system, which affords an advantage in that it is possible to obtain clear observation images having little noise, such as flare.
  • the glass members can be constructed using environmentally conscious materials that do not include harmful substances, such as lead, which is preferable from the viewpoint of environmental protection.
  • FIG. 1 shows the configuration of a fluorescence microscope according to an embodiment of the present invention.
  • FIG. 2 shows the configuration of a fluorescence filter set according to an embodiment of the present invention, which is used in the fluorescence microscope in FIG. 1 .
  • FIG. 3 schematically shows a transmission characteristic of a dichroic mirror according to an embodiment of the present invention, which is used in the fluorescence filter set in FIG. 2 , compared to the transmission characteristics of dichroic mirrors of the related art.
  • FIG. 4 schematically shows together transmission characteristics of first and second optical filters and a dichroic mirror constituting the fluorescence filter set in FIG. 2 .
  • FIG. 5 shows a transmission characteristic of the dichroic mirror in FIG. 3 , according to a first Example.
  • FIG. 6 shows a transmission characteristic of the dichroic mirror in FIG. 3 , according to a second Example.
  • FIG. 7 shows a transmission characteristic of the dichroic mirror in FIG. 3 , according to a third Example.
  • FIG. 8 shows a transmission characteristic of the dichroic mirror in FIG. 3 , according to a fourth Example.
  • FIG. 9 shows a transmission characteristic of the dichroic mirror in FIG. 3 , according to a fifth Example.
  • a dichroic mirror, a fluorescence filter set, and a fluoroscopy apparatus will be described below with reference to FIGS. 1 to 8 .
  • FIG. 1 The overall configuration of a fluoroscopy apparatus 1 according to this embodiment is shown in FIG. 1 .
  • the fluoroscopy apparatus 1 includes a light source 2 ; an illumination optical system 3 for efficiently guiding light L 1 emitted from the light source 2 ; a fluorescence filter set 4 to which the light L 1 from the illumination optical system is introduced and which emits excitation light L 2 in a direction perpendicular to the incident direction of the light L 1 , and in addition, which transmits in a straight line fluorescence L 3 returning from the direction in which the excitation light L 2 is emitted; an objective optical system 5 for focusing the emitted excitation light L 2 onto a specimen A; and an observation optical system 6 for observing the fluorescence L 3 produced in the specimen A and transmitted through the objective optical system 5 and the fluorescence filter set 4 .
  • the observation optical system 6 includes an imaging lens 7 and a detecting device 8 , for example, a CCD camera.
  • the fluorescence filter set 4 has a configuration in which a first optical filter (first wavelength-selecting member) 9 ; a dichroic mirror 10 ; and a second optical filter (second wavelength-selecting member) 11 are disposed inside a casing 12 .
  • the first optical filter 9 transmits only excitation light L 2 of a specific wavelength from the light L 1 incident thereon from the light source 2 .
  • the dichroic mirror 10 is disposed at an angle of 45° with respect to the incident direction so as to reflect the excitation light L 2 transmitted by the first optical filter 9 in a direction orthogonal to the incident direction thereof.
  • the second optical filter 11 selectively transmits the fluorescence L 3 transmitted by the dichroic mirror 10 .
  • the dichroic mirror 10 includes a dielectric multilayer film 10 b , formed by alternately laminating high-refractive-index layers and low-refractive-index layers on a planar transparent substrate 10 a.
  • the high-refractive-index layers are dielectric layers formed of a high-refractive-index material with a refractive index of 2.0 or more.
  • the low-refractive-index layers are dielectric layers formed of a low-refractive-index material with a refractive index of 1.5 or less.
  • the optical film thicknesses of these high-refractive-index and low-refractive-index layers are set to be from 1.5 ⁇ 0 /4 to 2.5 ⁇ 0 /4, with respect to a design wavelength ⁇ 0 .
  • the dielectric multilayer film 10 b preferably includes alternating layers formed by alternately laminating 50 to 150 high-refractive-index layers and low-refractive-index layers.
  • the design wavelength ⁇ 0 is a first-harmonic central wavelength at an incident angle of 0°.
  • the dichroic mirror 10 has a reflection band at the short wavelength side of the design wavelength ⁇ 0 . In other words, it does not use the first-harmonic wavelength band located close to the design wavelength ⁇ 0 as a reflection band B; rather, it uses a wavelength band of a second harmonic or higher, which appears at the short wavelength side of the design wavelength ⁇ 0 , as the reflection band B.
  • a dichroic mirror of the related art uses a wavelength band where the transmission characteristic is approximately 0%, which appears in the first-harmonic wavelength band located close to the design wavelength ⁇ 0 , as the reflection band B.
  • the reason for this is that a region where the transmission characteristic is approximately 0% in a wavelength band of a second high harmonic or higher, which appears at the short wavelength side of the design wavelength ⁇ 0 , is extremely narrow, and therefore, use of such a wavelength band as the reflection band B is completely out of the question.
  • the dichroic mirror 10 is formed by alternating 50 to 150 high-refractive-index layers and low-refractive-index layers, it is possible to ensure a sufficiently wide wavelength band where the transmission characteristic is approximately 0% in a wavelength band of this second harmonic or higher, and this band can thus be used as the reflection band B.
  • the dichroic mirror 10 has an ideal transmission characteristic, as schematically shown by the characteristic in FIG. 3 .
  • Comparative Example 1 in FIG. 3 is an example in which the optical film thicknesses of the high-refractive-index layer and the low-refractive-index layer are both ⁇ 0 /4
  • Comparative Examples 2 and 3 are examples in which the optical film thicknesses of the high-refractive-index layer and the low-refractive-index layer are ⁇ 0 /4 and 3 ⁇ 0 /4, respectively.
  • the number of alternately laminated layers in each case is about 20.
  • Comparative Example 1 a large step occurs in the transmission characteristic due to splitting of P-polarized and S-polarized light, as a result of disposing the dichroic mirror at an angle of 45° with respect to the incident optical axis of the excitation light L 2 .
  • Comparative Examples 2 and 3 have no step but they suffer from the drawback that in the reflection band B, the transmission characteristics rise gently from approximately 0%, and the transmission characteristics in a fluorescence-transmitting band C gradually reach about 100%. In particular, there is a problem with Comparative Example 3 in that the transmittance in the fluorescence-transmitting band C drops suddenly at longer wavelengths without reaching 100%.
  • the dichroic mirror 10 from the reflection band B where the transmittance is approximately 0%, exhibits a small step due to splitting of P-polarized and S-polarized light, and quickly reaches about 100% in the fluorescence-transmitting band C. Moreover, the fluorescence-transmitting band C extends to long wavelengths.
  • the dichroic mirror according to this embodiment it is possible to make the reflection band B and the fluorescence-transmitting band C close to each other. Since the wavelength band of the excitation light L 2 and the wavelength band of the fluorescence L 3 produced by that excitation light L 2 are close to each other, making the reflection band B and the fluorescence-transmitting band C close to each other allows the excitation light L 2 to be irradiated onto the specimen A in such a way that the fluorescence L 3 is efficiently produced in the specimen A.
  • the fluorescence filter set 4 which uses the dichroic mirror 10 , it is possible to set a transmission characteristic F 1 for the excitation light L 2 selected by the first optical filter 9 and a transmission characteristic F 2 for the fluorescence L 3 selected by the second optical filter 11 close to each other.
  • the transmission characteristic F 3 of the dichroic mirror 10 rises sharply from the reflection band B where the transmittance is approximately 0%, even if the transmission characteristic F 1 of the first optical filter 9 is set at comparatively longer wavelengths, the problem of the excitation light L 2 transmitted by the first optical filter 9 being transmitted by the dichroic mirror 10 can be prevented.
  • the excitation light L 2 is not transmitted by the dichroic mirror 10 , it is not scattered inside the casing 12 , and the excitation light L 2 can thus be prevented from passing through the second optical filter 11 without using colored glass having low angular dependency as the second optical filter 11 .
  • the transmission characteristic F 3 of the dichroic mirror 10 quickly reaches the fluorescence-transmitting band C where the transmittance is approximately 100%, even though the transmission characteristic F 2 of the second optical filter 11 is set at comparatively shorter wavelengths, part of the fluorescence L 3 that can pass through the second optical filter 11 can be prevented from being reflected at the dichroic mirror 10 , which allows the fluorescence L 3 to be efficiently collected.
  • the transmission characteristic F 3 of the dichroic mirror 10 has the fluorescence-transmitting band C which extends to long wavelengths, it is possible to construct a fluorescence filter set 4 that is capable of multiple uses, even when to collect long-wavelength fluorescence L 3 .
  • the glass components constituting the fluoroscopy apparatus 1 can be fabricated using only environmentally conscious materials that do not include harmful substances, such as lead and cadmium, which provides an advantage in that it is possible to construct a fluoroscopy apparatus 1 that is desirable from the viewpoint of environmental protection.
  • the optical film thicknesses of the dichroic multilayer film 10 b according to a first Example, used in the dichroic mirror 10 according to the embodiment described above, are shown in Table 1.
  • the transmission characteristic F 3 of the dichroic mirror 10 according to the first Example is shown in FIG. 5 .
  • the symbol H in Table 1 indicates a high-refractive-index layer, and the symbol L indicates a low-refractive index layer.
  • the film forming method used to fabricate the dichroic mirror may be ion-assisted deposition, RF ion plating, RF sputtering, ion-beam sputtering, and so on, so long as the same results are achieved.
  • the coefficients representing the optical film thickness of each layer are the same as the coefficients in Table 1. However, since value of the design wavelength ⁇ 0 is different, the optical film thicknesses are different from those in the first Example.
  • the coefficients representing the optical film thicknesses of each layer are the same as the coefficients in Table 1. However, since the value of the design wavelength ⁇ 0 is different, the optical film thicknesses are different from those in the first and second Examples.
  • the coefficients representing the optical film thicknesses of each layer are the same as the coefficients shown in Table 2. However, since the value of the design wavelength ⁇ 0 is different, the optical film thicknesses are different from those in the fourth Example.
  • dichroic mirror 10 having an ideal transmission characteristic F 3 which sharply rises from the reflection band B where the transmittance is approximately 0% to the fluorescence-transmitting band C where the transmittance is approximately 100%, which exhibits an extremely small step due to splitting of P-polarized and S-polarized light, and in which the fluorescence-transmitting band C extends to comparatively long wavelengths.
  • the fluorescence filter set 4 described above it is possible to use a laser light source instead of the first optical filter 9 .
  • possible wavelengths of the laser light source include, for example, 488 nm, 512 nm, 543 nm, and 633 nm.
  • the light from such a laser light source, serving as the excitation light L 2 is reflected by the dichroic mirror 10 and produces fluorescence L 3 when irradiating the specimen A.
  • excitation light L 2 having high wavelength purity can be obtained, which allows highly efficient fluoroscopy to be carried out.

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  • Physics & Mathematics (AREA)
  • General Physics & Mathematics (AREA)
  • Optics & Photonics (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
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  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Microscoopes, Condenser (AREA)
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  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
US11/184,814 2004-07-22 2005-07-20 Dichroic mirror, fluorescence filter set, and fluoroscopy apparatus Expired - Lifetime US7298550B2 (en)

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JP2004214620A JP4488299B2 (ja) 2004-07-22 2004-07-22 ダイクロイックミラー、蛍光フィルタセットおよび顕微鏡装置

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KR20110084328A (ko) * 2008-11-19 2011-07-21 쓰리엠 이노베이티브 프로퍼티즈 컴파니 고 내구성 색상 조합기
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US7825578B2 (en) * 2006-03-01 2010-11-02 Nichia Corporation Yellow light emitting device with high luminance
US20110032609A1 (en) * 2009-08-10 2011-02-10 Chroma Technology Corporation Microscope cube
US8488238B2 (en) * 2009-08-10 2013-07-16 Chroma Technology Corporation Microscope cube

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